Information on EC 4.2.1.66 - cyanide hydratase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
4.2.1.66
-
RECOMMENDED NAME
GeneOntology No.
cyanide hydratase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
formamide = cyanide + H2O
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
elimination
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
cyanide detoxification II
-
-
Cyanoamino acid metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
formamide hydro-lyase (cyanide-forming)
-
CAS REGISTRY NUMBER
COMMENTARY hide
37292-83-4
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Helminthosporium sorghicola
-
-
-
Manually annotated by BRENDA team
no activity in higher plants
-
-
-
Manually annotated by BRENDA team
Periconia circinata
-
-
-
Manually annotated by BRENDA team
Trichoderma spp.
several strains with varying levels of enzyme activity, constitutive expression of the gene encoding the enzyme
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-cyanopyridine + H2O
?
show the reaction diagram
acetonitrile + H2O
?
show the reaction diagram
-
-
?
acrylonitrile + H2O
?
show the reaction diagram
acrylonitrile + H2O
acrylamide
show the reaction diagram
benzonitrile + H2O
?
show the reaction diagram
best substrate for nitrilase activity
-
?
crotononitrile + H2O
?
show the reaction diagram
crotononitrile + H2O
crotonic acid amide
show the reaction diagram
cyanide + H2O
formamide
show the reaction diagram
fumaronitrile + H2O
?
show the reaction diagram
HCN + H2O
formamide
show the reaction diagram
methacrylonitrile + H2O
2-methylprop-2-enamide
show the reaction diagram
methacrylonitrile + H2O
?
show the reaction diagram
propionitrile + H2O
?
show the reaction diagram
-
-
?
tetracyanonickelate II + H2O
?
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
cyanide + H2O
formamide
show the reaction diagram
HCN + H2O
formamide
show the reaction diagram
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
CN-
-
rapid inactivation above 150 mM
p-chloromercuribenzoate
-
-
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
cyanide
additional information
Trichoderma spp.
-
no induction of enzyme expression by cyanide in the different strains
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3.7
2-Cyanopyridine
-
at pH 8.0 and 30C
15 - 43
CN-
109
cyanide
-
at pH 8.0 and 30C
14.7
Fumaronitrile
-
at pH 8.0 and 30C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.3 - 5.7
-
-
6.5 - 7.5
-
immobilized enzyme
9 - 10.7
-
-
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.2 - 9
5.5 - 10.5
-
-
6 - 8.5
additional information
-
broad range
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
27 - 50
-
incubated at 27C, 37C, 43C, and 50C, activity measured at time points ranging between 0 and 48 h, at 37C and 43C most stable with no loss of activity after 48 h at 37C, about 40% of activity remaining after 48 h at 43C, at 50C activity below 5% by 48 h
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40900
-
x * 40900, calculation from sequence of cDNA
43000
-
x * 43000, SDS-PAGE
45000
-
x * 45000, SDS-PAGE
300000 - 1000000
-
gel filtration
300000 - 400000
-
gel filtration
300000
additional information
-
the helical structure of cyanide hydratase is determined at a resolution of about 33 A by applying the iterative helical real-space reconstruction method algorithm to freeze-dried unidirectionally shadowed filaments. These filaments are left-handed
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oligomer
polymer
tetramer
-
n * 40000
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
side-chain modification
-
N-glycosylation possibly at residue 286-288
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 10
-
unstable below pH 5.5
730851
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22 - 24
-
the activity is totally lost by incubation of the mycelia for 16 h at 22-24C
25
-
50% loss of activity in 2 days
27 - 50
28 - 55
-
the enzyme shows a half-life of 5.6 hat 28C and is unstable at temperatures above 35C. The enzyme is still active at 50C with ca. 36% of its maximum activity, but nearly inactive at 55C
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80C, more than 2 years
-
4C, 4 days, loss of 50% activity
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4C, 5-10% loss of activity per week
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Hi-Prep 16/10 Q FF column chromatography and Sephacryl S-200 gel filtration
-
recombinant protein, gel filtration
recombinant protein, gel filtration and SDS-PAGE
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA sequence analysis
expressed in Escherichia coli BL21-Gold(DE3) cells
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expressed in Escherichia coli, pET26b construct p3460, BL21(lambda-DE3) pLysS cells
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expressed in Escherichia coli, strains and plasmids listed
expression in Escherichia coli
-
expression in Escherichia coli BL21
-
overexpression of the wild-type and mutant enzymes in Escherichia coli XL1-Blue
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D275E
-
site-directed mutagenesis, 0-29% reduced activity compared to the wild-type enzyme
F170L
-
site-directed mutagenesis, inactive mutant compared to the wild-type enzyme
K136R
-
site-directed mutagenesis, 52% reduced activity compared to the wild-type enzyme
M302S
-
site-directed mutagenesis, 22% reduced activity compared to the wild-type enzyme
S13A
-
site-directed mutagenesis, 19-47% reduced activity compared to the wild-type enzyme
T12P
-
site-directed mutagenesis, inactive mutant
T12Q
-
site-directed mutagenesis, 10-48% reduced activity compared to the wild-type enzyme
V281A
-
site-directed mutagenesis, 8% reduced activity compared to the wild-type enzyme
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
-
optical biotest for determination of cyanide traces in marine fish using microbial cyanide hydratase and formate dehydrogenase
biotechnology
additional information
Show AA Sequence (185 entries)
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